LOCUS       sequence-364987-        6091 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-1 vector encoding PhCMV-driven red fluorescent protein mRuby2 
            localized to the ER (PhCMV-Igk-mRuby2-KDEL-pA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6091)
  AUTHORS   Haellman V, Strittmatter T, Bertschi A, Stucheli P, Fussenegger M
  TITLE     A versatile plasmid architecture for mammalian synthetic biology 
            (VAMSyB).
  JOURNAL   Metab Eng. 2021 Jul;66:41-50. doi: 10.1016/j.ymben.2021.04.003. Epub
            2021 Apr 18.
  PUBMED    33857582
REFERENCE   2  (bases 1 to 6091)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Metab 
            Eng."; date: "2021-07"; volume: "66:41-50. doi"; pages: " 
            10.1016/j.ymben.2021.04.003. Epub 2021 Apr 18"
COMMENT     Sequence Label: pTS2395-Tier1-PhCMV-Igk_mRuby2_KDEL
FEATURES             Location/Qualifiers
     source          1..6091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        109..488
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        489..692
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     643..663
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     737..756
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        737..755
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     796..852
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="ETDTLLLWVLLLWVPGSTG"
     CDS             874..1581
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTHM
                     ALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAVA
                     KFAGLGGGMDELYK"
     CDS             1609..1638
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1751..1768)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1757..1981
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2027..2455
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2114..2133)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2324..2345
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2464..2484)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2469..2798
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2649..2784
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2711..2730
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2865..3659
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2919..2938)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3529..3548
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3833..3954
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3870..3889)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3924..3943
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4003..4019)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4003..4019)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4016..4038)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4027..4043
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4051..4057,4058..4075,4076..4081))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4096..4117
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4234..4251)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4405..4993)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4485..4504)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5164..5955,5956..6024))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5787..5806
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6025..6091,1..38))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
       61 cctgacgtcg acggatcggg agatctccac gcgtggtacc ctcgagttga cattgattat
      121 tgagtagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
      181 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      241 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      301 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      361 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      421 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      481 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      541 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      601 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      661 gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt
      721 actggcttat cgaaattaat acgactcact atagggagac ccaagctgtt cgaagcggaa
      781 ttcaccatga ctagtgagac agacacactc ctgctatggg tactgctgct ctgggttcca
      841 ggttccactg gtgacgctag tggtggttct ggtgtgtcta agggcgaaga gctgatcaag
      901 gaaaatatgc gtatgaaggt ggtcatggaa ggttcggtca acggccacca attcaaatgc
      961 acaggtgaag gagaaggcaa tccgtacatg ggaactcaaa ccatgaggat caaagtcatc
     1021 gagggaggac ccctgccatt tgcctttgac attcttgcca cgtcgttcat gtatggcagc
     1081 cgtactttta tcaagtaccc gaaaggcatt cctgatttct ttaaacagtc ctttcctgag
     1141 ggttttactt gggaaagagt tacgagatac gaagatggtg gagtcgtcac cgtcatgcag
     1201 gacaccagcc ttgaggatgg ctgtctcgtt taccacgtcc aagtcagagg ggtaaacttt
     1261 ccctccaatg gtcccgtgat gcagaagaag accaagggtt gggagcctaa tacagagatg
     1321 atgtatccag cagatggtgg tctgagggga tacactcata tggcactgaa agttgatggt
     1381 ggtggccatc tgtcttgctc tttcgtaaca acttacaggt caaaaaagac cgtcgggaac
     1441 atcaagatgc ccggtatcca tgccgttgat caccgcctgg aaaggttaga ggaaagtgac
     1501 aatgaaatgt tcgtagtaca acgcgaacac gcagttgcca agttcgccgg gcttggtggt
     1561 gggatggacg agctgtacaa ggctagcgga tccaaggacg agctgtaaga acaaaaactt
     1621 atttctgaag aggatctgtg atgacgtaac aattacgggt ctaccatcga gggcctgctc
     1681 gatctcccgg acgacgacgc cccgaagagg cggggtctag aaagctttga ggccggcctg
     1741 cagggatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     1801 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     1861 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     1921 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     1981 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     2041 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2101 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2161 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2221 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2281 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2341 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2401 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     2461 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2521 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     2581 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     2641 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2701 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     2761 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     2821 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     2881 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     2941 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3001 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3061 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3121 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3181 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3241 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3301 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3361 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     3421 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     3481 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     3541 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     3601 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     3661 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     3721 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     3781 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     3841 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3901 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     3961 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     4021 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4081 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4141 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4201 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4261 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4321 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4381 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4441 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4501 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4561 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4621 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4681 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4741 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4801 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4861 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4921 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4981 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5041 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5101 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5161 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5221 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5281 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5341 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5401 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5461 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5521 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5581 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5641 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5701 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5761 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5821 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5881 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5941 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6001 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6061 gggttattgt ctcatgagcg gatacatatt t
//
